Quantification of Type I Collagen α1 in Culture Medium Using Nano-Liquid Chromatography Tandem Mass Spectrometry

DOI DOI 機関リポジトリ
  • GONG Ao
    Department of Biosignaling and Radioisotope Experiment, Interdisciplinary Center for Science Research, Organization for Research and Academic Information, Shimane University, Izumo, Shimane 693-8501, Japan Department of Legal Medicine, Faculty of Medicine, Shimane University, Izumo, Shimane 693-8501, Japan School of Basic Medical Science, Ningxia Medical University, Yinchuan, Ningxia 750004, China
  • YAMADA Kazuo
    Department of Biosignaling and Radioisotope Experiment, Interdisciplinary Center for Science Research, Organization for Research and Academic Information, Shimane University, Izumo, Shimane 693-8501, Japan Department of Legal Medicine, Faculty of Medicine, Shimane University, Izumo, Shimane 693-8501, Japan
  • JING Li
    School of Basic Medical Science, Ningxia Medical University, Yinchuan, Ningxia 750004, China
  • TAKESHITA Haruo
    Department of Legal Medicine, Faculty of Medicine, Shimane University, Izumo, Shimane 693-8501, Japan
  • MATSUMOTO Ken-ichi
    Department of Biosignaling and Radioisotope Experiment, Interdisciplinary Center for Science Research, Organization for Research and Academic Information, Shimane University, Izumo, Shimane 693-8501, Japan

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抄録

Type I collagen, which consists of two α1 and one α2 chains, is an abundant extracellular matrix (ECM) protein. Changes in type I collagen are often associated with various diseases including fibrosis, osteogenesis imperfecta (OI), and Ehlers-Danlos syndrome (EDS). In the present study, we developed a method for quantification of type I collagen α1 (COL1A1) in a culture medium of LX-2 human hepatic stellate cells by using nano-liquid chromatography tandem mass spectrometry (nano-LC/MS/MS). After selecting a specific peptide of COL1A1, unlabeled and stable isotope-labeled peptides were used for quantitative analysis of COL1A1 by nano-LC/MS/MS. The concentration of secreted COL1A1 in the LX-2 cell culture medium was 38.78 ng/mL. The results indicate that this method is useful for quantifying COL1A1 in a cell culture medium.

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